Pharmaceutically active morpholinol

ABSTRACT

Disclosed is the compound (+)-(2S,3S)-2-(3-chlorophenyl)-3,5,5-trimethyl-2-morpholinol and pharmaceutically acceptable salts and solvates thereof, pharmaceutical compositions comprising them, and processes for their preparation; also disclosed is a method of treating depression, attention deficit hyperactivity disorder (ADHD), obesity, migraine, pain, sexual dysfunction, Parkinson&#39;s disease, Alzheimer&#39;s disease, addiction to cocaine or tobacco products, seasonal affective disorder, chronic fatigue, narcolepsy or cognitive impairment using such compound, salts, solvates or compositions.

[0001] This application is a continuation-in-part of U.S. Ser. No.09/886,391 filed Jun. 22, 2001, which is a divisional application ofU.S. Ser. No. 09/233,531, filed Jan. 20, 1999, now U.S. Pat. No.6,274,579.

[0002] This invention relates to an optically pure morpholinol, saltsand solvates thereof, pharmaceutical formulations containing them andprocesses for their preparation and use.

BACKGROUND OF THE INVENTION

[0003] Bupropion hydrochloride,(±)-1-(3-chlorophenyl-2-[(1,1-dimethylethyl)-amino]-1-propanonehydrochloride, is the active ingredient of Wellbutrin® which is marketedin the United States for the treatment of depression. It is also theactive ingredient of Zyban® which is marketed in the United States as anaid to smoking cessation. Bupropion is a relatively weak inhibitor ofthe neuronal uptake of noradrenaline (NA), serotonin and dopamine (DA),and does not inhibit monoamine oxidase. While the mechanism of action ofbupropion, as with other antidepressants, is unknown, it is presumedthat this action is mediated by noradrenergic and/or dopaminergicmechanisms. Available evidence suggests that Wellbutrin® is a selectiveinhibitor of noradrenaline (NA) at doses that are predictive ofantidepressant activity in animal models. See Ascher, J. A., et al.,Bupropion: A Review of its Mechanism of Antidepressant Activity. Journalof Clinical Psychiatry, 56: p. 395-401,1995.

[0004] Bupropion is extensively metabolized in man as well as laboratoryanimals. Urinary and plasma metabolites include biotransformationproducts formed via hydroxylation of the tert-butyl group and/orreduction of the carbonyl group of bupropion. Four basic metaboliteshave been identified. They are the erythro- and threo-amino alcohols ofbupropion, the erythro-amino diol of bupropion, and a morpholinolmetabolite. These metabolites of bupropion are pharmacologically active,but their potency and toxicity relative to bupropion have not been fullycharacterized. Because the plasma concentrations of the metabolites arehigher than those of bupropion, they may be of clinical importance.

[0005] The morpholinol metabolite(+/−)-(2R*,3R*)-2-(3-chlorophenyl)-3,5,5-trimethyl-2-morpholinolhydrochloride is believed to be formed from hydroxylation of thetert-butyl group of bupropion.

Morpholinol Metabolite of Bupropion HCl SUMMARY OF THE INVENTION

[0006] It has now surprisingly been discovered that despite the (−) formof the morpholinol metabolite predominating significantly in humanplasma samples, it is the (+) enantiomer,(+)-(2S,3S)-2-(3-chlorophenyl)-3,5,5-trimethyl-2-morpholinol in whichthe activity resides.

[0007] Thus the present invention provides, in one aspect, the compoundof formula (I),(+)-(2S,3S)-2-(3-chlorophenyl)-3,5,5-trimethyl-2-morpholinol orpharmaceutically acceptable salts and solvates thereof.

[0008] Another aspect of the invention is pharmaceutical compositionscomprising a compound of formula (I) or pharmaceutically acceptablesalts and solvates thereof together with one or more pharmaceuticallyacceptable carriers, diluents or excipients.

[0009] A further aspect of the present invention is the use of acompound of formula (I) or pharmaceutically acceptable salts andsolvates thereof in therapy.

[0010] Yet another aspect of the invention provides methods of treatingdepression, attention deficit hyperactivity disorder (ADHD), obesity,migraine, pain, sexual dysfunction, Parkinson's disease, Alzheimer'sdisease, or addiction to cocaine or tobacco products in a mammal (humanor animal subject) comprising the administration to said subject of aneffective amount of a compound of formula (I) or pharmaceuticallyacceptable salts and solvates thereof or pharmaceutical compositionsthereof.

[0011] Yet another aspect of the present invention is the use of thecompound of formula (I) or pharmaceutically acceptable salts andsolvates thereof or pharmaceutical compositions thereof in thepreparation of a medicament for the treatment of depression, attentiondeficit hyperactivity disorder (ADHD), obesity, migraine, pain, sexualdysfunction, Parkinson's disease, Alzheimer's disease, addiction tococaine or tobacco products.

[0012] Yet another aspect of the present invention provides a method oftreatment of seasonal affective disorder in a mammal (human or animalsubject) comprising the administration to said subject of an effectiveamount of a compound of formula (I) or pharmaceutically acceptable saltsand solvates thereof or pharmaceutical compositions thereof.

[0013] Yet another aspect of the present invention provides a method oftreatment of chronic fatigue in a mammal (human or animal subject)comprising the administration to said subject of an effective amount ofa compound of formula (I) or pharmaceutically acceptable salts andsolvates thereof or pharmaceutical compositions thereof.

[0014] Yet another aspect of the present invention provides a method oftreatment of narcolepsy in a mammal (human or animal subject) comprisingthe administration to said subject of an effective amount of a compoundof formula (I) or pharmaceutically acceptable salts and solvates thereofor pharmaceutical compositions thereof.

[0015] Yet another aspect of the present invention provides a method oftreatment of cognitive impairment in a mammal (human or animal subject)comprising the administration to said subject of an effective amount ofa compound of formula (I) or pharmaceutically acceptable salts andsolvates thereof or pharmaceutical compositions thereof.

DESCRIPTION OF THE DRAWINGS

[0016]FIG. 1. Effect of Compounds at 25 mg/kg (ip) on TBZ-InducedDepression

[0017]FIG. 2. Dose Response of Compound of Formula I Against TBZ-InducedDepression (Compounds administered 30 minutes prior to TBZ, Male, CD-1Mice, i.p., n=6)

[0018]FIG. 3. Dose Response of Compound of Formula II AgainstTBZ-Induced Depression (Compounds administered 30 minutes prior to TBZ,Male, CD-1 Mice, i.p., n=6)

DETAILED DESCRIPTION OF THE INVENTION

[0019] It will be appreciated that references herein to “treatment”extend to prophylaxis, prevention of recurrence and suppression oramelioration of symptoms (whether mild, moderate or severe) as well asthe treatment of established conditions.

[0020] The term “treatment of seasonal affective disorder” includes, butis not limited to, the treatment of seasonal affective disorder (SAD),major depressive disorder with seasonal pattern, and sub-syndromal SAD(‘winter blues’) and encompasses the prevention and alleviation, bothpartial and complete, of one or more of the symptoms thereof. SAD is,for many sufferers, a seriously disabling illness, preventing them fromfunctioning normally without continuous medical treatment. Sub-syndromalSAD, although a milder form of the condition, can still be debilitating.The symptoms of SAD usually recur regularly each fall or winter and caninclude a number of the following: change in appetite, change in weight,sleep problems (including a desire to oversleep, difficulty stayingawake, disturbed sleep, early morning wakening), lethargy, decreasedenergy level, leaden paralysis, difficulty concentrating, depression(feelings of misery, guilt, loss of self-esteem, hopelessness, apathy),social interaction problems (irritability, desire to avoid socialcontact, increased fear of social rejection), anxiety (tension,inability to tolerate stress), loss of libido, and mood changes.

[0021] The term “treatment of chronic fatigue” includes, but is notlimited to, treatment of persistent (≧6 months) fatigue that results ina significant reduction in activity and/or daily function for thesufferer. Such fatigue is often made worse by exertion, exercise, andother stresses and may be accompanied by other symptoms including sorethroat, painful lymph nodes, muscle discomfort (pain), prolonged fatigueafter exercise, generalised headaches, migratory arthralgias,neuropsychiatric complaints (impaired memory or concentration, abnormalvisual tolerance of light, loss of vision within the normal field ofvision, forgetfulness, irritability, confusion, depression) and sleepdisorders. Encompassed within the term is the treatment of symptoms ofchronic fatigue syndrome (with or without co-morbid non-melancholic ornon-psychotic mood disorders), chronic fatigue associated withdepression and other mood disorders, idiopathic chronic fatigue, andchronic fatigue associated with other conditions such as multiplesclerosis, organic brain disorders, and fibromyalgia. A preferredcondition for treatment is chronic fatigue syndrome.

[0022] The term “treatment of narcolepsy” includes, but is not limitedto, the treatment of excessive sleepiness, including in the form ofsleep attacks, short (usually lasting a hour or less) episodes of aseemingly irresistible need to sleep, and disrupted nocturnal sleep.Other symptoms include brief periods of loss of muscle tone (cataplexy)often in response to a sudden emotional reaction and/or recurrentintrusions of elements of rapid eye movement (REM) sleep into thetransition between sleep and wakefulness, as manifested by eitherhypnopompic or hypnagogic hallucinations or sleep paralysis at thebeginning or end of sleep episodes.

[0023] The term “treatment of cognitive impairment” includes, but is notlimited to, the treatment of impairment of cognitive functions(including attention, orientation, learning, memory consolidation andrecall, reaction time, and language function). Such impairment may be asa result of a number of causes, including ADHD, stroke, Alzheimer'sdisease or other dementia states, as well as other acute or sub-acuteconditions that may cause cognitive decline such as delirium ordepression (pseudodementia states).

Preparation

[0024] The compound of formula (I) or pharmaceutically acceptable saltsand solvates thereof may be prepared by first synthesizing the racemateof the morpholinol metabolite of bupropion and subsequently separatingthe (+) and (−) enantiomers of the racemate via HPLC.

[0025] The racemate of the morpholinol metabolite of bupropionhydrochloride((+/−)-(2R*,3R*)-2-(3-chlorophenyl)-3,5,5-trimethyl-2-morpholinolhydrochloride) may be synthesized by the following process. To3′-chloropropiophenone (10.0 g, 0.06 mol) in dioxane (50 mL) was added asolution of dioxane dibromide (14.9 g, 0.06 mol) in dioxane (50 mL). Thereaction mixture was stirred for 2 h at ambient temperature and pouredinto a mixture of ice and water (500 mL). The mixture was extractedseveral times with methylene chloride. The combined extracts were dried(Na₂SO₄) and concentrated in vacuo to give 14.8 g (85%) of2-bromo-3′-chloropropiophenone as a pale yellow oil. This was usedwithout further purification. NMR (300 Mhz, CDCl₃); δ 7.99 (m, 1H), 7.90(d, 1H), 7.57 (d, 1H), 7.44 (t, 1H), 5.22 (q, 1H), 1.91 (t, 3H).

[0026] To a solution of 2-bromo-3′-chloropropiophenone (19.3 g, 0.08mol) in MeOH (100 mL) was added dropwise a solution of2-amino-2-methyl-1-propanol (27.8 g, 0.31 mol) in methanol (200 mL) atambient temperature. The mixture was stirred for 18 h and concentratedin vacuo. The residue was partitioned between water and diethyl ether.The combined organic phase was extracted with 10% aqueous hydrogenchloride. The combined aqueous acid extracts were chilled in an ice bathand made basic with 40% aqueous sodium hydroxide. The mixture wasextracted with diethyl ether, the combined diethyl ether extracts werewashed with water and saturated sodium chloride solution, dried (K₂CO₃)and concentrated in vacuo to give 15.0 g (75%) of(+/−)-(2R*,3R*)-2-(3-chlorophenyl)-3,5,5-trimethyl-2-morpholinol as anoff-white solid.

[0027] (+/−)-(2R*,3R*)-2-(3-chlorophenyl)-3,5,5-trimethyl-2-morpholinolmay be converted to(+/−)-(2R*,3R*)-2-(3-chlorophenyl)-3,5,5-trimethyl-2-morpholinolhydrochloride by the following process. A 6.0 g sample was dissolved indiethyl ether, chilled in an ice bath and ethereal hydrogen chlorideadded until the mixture was acidic. The resulting solid was filtered andrecrystallized from ethanol/diethyl ether/ethereal hydrogen chloridemixtures to give 4.93 g of(+/−)-(2R*,3R*)-2-(3-chlorophenyl)-3,5,5-trimethyl-2-morpholinolhydrochloride as a white solid: m.p. 202-203° C. NMR (80 Mhz, DMSO-d₆);δ 10.9 (br, 1H), 8.85 (br, 1H), 7.60-7.41 (m, 5H), 4.04 (d, 1H), 3.50(d, 1H), 3.37 (br s, 1H), 1.58 (s, 3H), 1.34 (s, 3H), 1.03 (d, 3H).Anal. Calcd for C₁₃H₁₉Cl₂NO₂: C, 53.43; H, 6.55; N, 4.79. Found: C,53.54; H, 6.58; N, 4.75.

[0028] (+/−)-(2R*,3R*)-2-(3-chlorophenyl)-3,5,5-trimethyl-2-morpholinolhydrochloride may be converted back to its free base by the followingprocess. A 3.0 g sample of(+/−)-(2R*,3R*)-2-(3-chlorophenyl)-3,5,5-trimethyl-2-morpholinolhydrochloride was dissolved in water (100 mL) and diethyl ether wasadded (200 mL). The mixture was chilled in an ice bath and the pH wasadjusted to >10 with 1.0N aqueous sodium hydroxide. After stirring for30 min., the phases were separated and the aqueous phase was extractedwith diethyl ether. The combined diethyl ether extracts were dried(Na₂SO₄) and concentrated in vacuo to give 2.6 g of(+/−)-(2R*,3R*)-2-(3-chlorophenyl) -3,5,5-trimethyl-2-morpholinol as awhite solid. This was used without further purification for the chiralchromatography described below.

[0029] The (+) and (−) enantiomers of(+/−)-(2R*,3R*)-2-(3-chlorophenyl)-3,5,5-trimethyl-2-morpholinol may beseparated by the following process.(+/−)-(2R*,3R*)-2-(3-chlorophenyl)-3,5,5-trimethyl-2-morpholinol (2.54gms.) was dissolved in 250 ml of 2:8 Isopropyl alcohol:Hexane (both HPLCgrade). A Daicel Chiralcel OD column (2×25 cm.) was equilibrated for onehour at 8 ml./min.in the elution solvent, 1:9:0.2Isopropanol:Hexane:Diethylamine. The solution of the(+/−)-(2R*,3R*)-2-(3-chlorophenyl)-3,5,5-trimethyl-2-morpholinol wasinjected in 1 ml. aliquots by an automated Waters Prep LC 2000, using aWaters 510 EF pump for injections. Each run was 15 minutes in length,using the conditions listed before. The separated optical isomers werecollected by fraction collector (Waters) at a 2% above baselinethreshold, based on 2 absorbance units full scale at 240 nm (Waters 490EUV detector). Each optical isomer solution was evaporated on a rotaryevaporator at 40 degrees Centigrade and aspirator vacuum. After dryingfor 6 hours under high vacuum at room temperature, optical isomer 1weighed 1.25 gm. and optical isomer 2 weighed 1.26 gm.

[0030] The enantiomeric purity of each isomer was assayed by analyticalchiral HPLC on a Waters 860 HPLC with 996 Photodiode Array detector,using a Daicel Chiralcel OD-H column (4.6×250 mm.) eluted with 1:9:0.2Isopropyl alcohol: Hexane:Diethylamine at 1 ml/min. Optical isomer 1 was100% pure (R.T. 6.117 min.). Optical isomer 2 was 99.19% pure (R.T.6.800 min.), containing 0.81% optical isomer 1 (R.T. 6.133 min.).

[0031] Hydrochloride salts of the separated enantiomers were obtained bythe following processes. 1.25 g (0.005 mol) of optical isomer 1(retention time 6.117 min)((−)-(2R,3R)-2-(3-chlorophenyl)-3,5,5-trimethyl-2-morpholinol) wasdissolved in diethyl ether. The solution was filtered and the filtratewas chilled in an ice-bath adding ethereal hydrogen chloride until thesolution was acidic. After standing at ambient temperature for 24 h, theresulting solid was filtered, washed with diethyl ether and dried in avacuum oven at 60° C. for 18 h to give 1.32 g (90%) of(−)-(2R,3R)-2-(3-chlorophenyl)-3,5,5-trimethyl-2-morpholinolhydrochloride as a white solid: mp 208-209° C. NMR (300 Mhz, DMSO-d₆); δ9.72 (br, 1H), 8.76 (br, 1H), 7.54-7.41 (m, 5H), 3.98 (d, 1H), 3.52 (d,1H), 3.37 (br s, 1H), 1.53 (s, 3H), 1.29 (s, 3H), 0.97 (d, 3H). Anal.Calcd for C₁₃H₁₉Cl₂NO₂: C, 53.43; H, 6.55; N, 4.79. Found: C, 53.35; H,6.57; N, 4.71. (α)_(D) ^(20° C.)=−33.2° (0.67, 95% EtOH)

[0032] 1.26 g (0.005 mol) of optical isomer 2 (retention time 6.800 min)(+)-(2S,3S)-2-(3-chlorophenyl)-3,5,5-trimethyl-2-morpholinol) wasdissolved in diethyl ether. The solution was filtered and the filtratewas chilled in an ice-bath adding ethereal hydrogen chloride until thesolution was acidic. After standing at ambient temperature for 24 h, theresulting solid was filtered, washed with diethyl ether and dried in avacuum oven at 60° C. for 18 h to give 1.36 g (93%) of(+)-(2S,3S)-2-(3-chlorophenyl)-3,5,5-trimethyl-2-morpholinolhydrochloride as a white solid: mp 208-209° C. NMR (300 Mhz, DMSO-d₆); δ9.87 (br, 1H), 8.76 (br, 1H), 7.54-7.41 (m, 5H), 3.99 (d, 1H), 3.51 (d,1H), 3.37 (br s, 1H), 1.54 (s, 3H), 1.30 (s, 3H), 0.98 (d, 3H). Anal.Calcd for C₁₃H₁₉Cl₂NO₂: C, 53.43; H, 6.55; N, 4.79. Found: C, 53.51; H,6.58; N, 4.73. (α)_(D) ^(20° C.)=+31.9° (0.64, 95% EtOH)

[0033] The absolute configuration of(+)-(2S,3S)-2-(3-chlorophenyl)-3,5,5-trimethyl-2-morpholinol wasdetermined by the following x-ray crystallographic method. Crystal Data:C₁₃H₁₈Cl₂NO₂, M=291, Orthorhombic, space group P2_(l)2_(l)2_(l),a=8.7348 (6), b=14.9824 (10), c=23.1605 (15) Å, V=3031 (4) Å³, Z=8,Dc=1.276 Mgm⁻³, F(000)=1226.95. Of 12224 reflections measured. 3764 wereunique and 2318 which had I>3.0σ(I) were used in subsequentcalculations. Data was collected on a Siemens SMART diffractometer usingomega scans and monochromated MoKα radiation (λ=0.71073 Å). Thepositions of all non-hydrogen atoms were determined by direct methodsand refined anisotropically. The hydrogen positions were all located indifference syntheses and included in subsequent refinement cycles usinga riding model and an idealized bond length of 0.96 Å. The absoluteconfiguration was determined by refinement of the Rogers' parameter andconfirmed by an analysis of the 185 best Bijvoet intensity differenceswhich indicated a probability of 0.006 that the model was in error.Least squares refinement minimized Σw(ΔF)² with weights based on counterstatistics. The final agreement factors were R_(f)=0.064 (0.108 for alldata), R_(w)=0.068 (0.081 for all data), and GoF=1.93. Referencesincluded E. J. Gabe, Y. Le Page, J.-P. Charland, F. L. Lee and P. S.White, Journal of Applied Crystallography, 22, 384-387 (1989) and D.Rogers, Acta Crystallographica, A37, 734-741, 1981.

Dosage and Formulation

[0034] The amount of compound of formula (I) required to achieve thedesired therapeutic effect will, of course depend on a number offactors, for example, the mode of administration, the recipient and thecondition being treated. In general, the daily dose will be in the rangeof 0.02 to 5.0 mg/kg. More particular ranges include 0.02 to 2.5 mg/kg,0.02 to 1.0 mg/kg, 0.02 to 0.25 mg/kg, 0.02 to 0.15 mg/kg and 0.02 to0.07 mg/kg.

[0035] The compound of formula (I) may be employed in the treatment ofdepression, attention deficit hyperactivity disorder (ADHD), obesity,migraine, pain, sexual dysfunction, Parkinson's disease, Alzheimer'sdisease, addiction to cocaine or tobacco products, seasonal affectivedisorder, chronic fatigue, narcolepsy or cognitive impairment as thecompound per se, but is preferably presented with one or morepharmaceutically acceptable carriers, diluents or excipients in the formof a pharmaceutical formulation. The carriers, diluents and exipientsmust, of course, be acceptable in the sense of being compatible with theother ingredients of the formulation and must not be deleterious to therecipient. The carrier may be a solid or a liquid, or both, and ispreferably formulated with the agent as a unit-dose formulation, forexample, a tablet.

[0036] The formulations include those suitable for oral, rectal,topical, buccal (e.g. sub-lingual) and parenteral (e.g. subcutaneous,intramuscular, intradermal or intravenous) administration.

[0037] Formulations suitable for buccal (sub-lingual) administrationinclude lozenges comprising a compound of formula (I) in a flavouredbase, usually sucrose and acacia or tragacanth, and pastilles comprisingthe agent in an inert base such as gelatin and glycerin or sucrose andacacia.

[0038] Formulations of the present invention suitable for parenteraladministration conveniently comprise sterile aqueous preparations of acompound of formula (I), preferably isotonic with the blood of theintended recipient. These preparations are preferably administeredintravenously, although administration may also be effected by means ofsubcutaneous, intramuscular, or intradermal injection. Such preparationsmay conveniently be prepared by admixing the agent with water andrendering the resulting solution sterile and isotonic with the blood.

[0039] Formulations suitable for rectal administration are preferablypresented as unit-dose suppositories. These may be prepared by admixinga compound of formula (I) with one or more conventional solid carriers,for example, cocoa butter, and then shaping the resulting mixture.

[0040] Formulations suitable for topical application to the skinpreferably take the form of an ointment, cream, lotion, paste, gel,spray, transdermal patch, aerosol, or oil. Carriers which may be usedinclude vaseline, lanolin, polyethylene glycols, alcohols, andcombinations of two or more thereof.

[0041] It should be understood that in addition to the ingredientsparticularly mentioned above, the formulations may include other agentsconventional in the art having regard to the type of formulation inquestion.

Biological Activity

[0042] Biological activity of the compound of formula (I) wasdemonstrated by in vitro uptake models and the tetrabenazine-inducedbehavioural depression model. The racemic morpholinol metabolite,(+/−)-(2R*,3R*)-2-(3-chlorophenyl)-3,5,5-trimethyl-2-morpholinol, isreferred to herein as “Racemate”. The (−) form of the morpholinolmetabolite is (−)-(2R,3R)-2-(3-chlorophenyl)-3,5,5-trimethyl-2-morpholinol or pharmaceuticallyacceptable salts and solvates thereof and is referred to herein as acompound of formula (II):

[0043] In vitro Synaptosomal Uptake Experiments

[0044] In vitro uptake was determined, as reported previously, usingsynaptosomes prepared from rat caudoputamen (for dopamine uptake) andhypothalamus (for NA and serotonin uptake) using [³H]-dopamine, [³H]-NAand [³H]-serotonin as transport substrates, respectively. See Eckhardt,S. B., R. A. Maxwell, and R. M. Ferris, A Structure-Activity Study ofthe Transport Sites for the Hypothalamic and Striatal CatecholamineUptake Systems. Similarities and differences. Molecular Pharmacology,21: p. 374-9,1982.

[0045] Synaptosomes for use in obtaining in vitro uptake data wereprepared from hypothalamus or striatum by gently homogenizing the tissuein a 0.3 M sucrose/25 mM Tris pH 7.4 buffer containing iproniazidphosphate to inhibit monoamine oxidase. The homogenate was centrifugedat 1100× g at 4° C. for 10 min and the supernatant was used for uptakestudies. The supernatant (˜1 mg tissue protein) was incubated with Kmconcentrations of [³H]-noradrenaline, [³H]-dopamine or [³H]-serotonin at37° C. for 5 minutes in Modified Krebs-Henseleit buffer (118 mM NaCl, 5mM KCl, 25 mM NaHCO₃, 1.2 mM NaH₂PO₄, 1.2 mM MgSO₄, 11 mM Dextrose, 2.5mM CaCl₂) in the absence and presence of drug. Under these conditionsuptake was linear with respect to both for substrate and tissue (with<5% total substrate transported). Non-specific uptake was defined asuptake at 0° C. [³H]-substrate, which had been transported intosynaptosomes, was separated from free [³H]-substrate by filtration overGF/B filters and washing with cold Krebs-Henseleit buffer. The filterswere counted for tritum in a liquid scintillation spectrometer.

[0046] The data for in vitro synaptosomal uptake are presented asTable 1. Among the 2 enantiomers of the morpholinol metabolite ofbupropion, the (+) enantiomer, the compound of formula (I), inhibitednoradrenaline (NA) uptake with an IC₅₀ of 2.2 μM. In contrast, the (−)enantiomer was ineffective at a concentration of 30 μM. On dopamine (DA)uptake, the compound of formula (I) had an IC₅₀ of ˜10 μM while the (−)enantiomer was inactive at 30 μM. Neither compound inhibited serotoninuptake at 30 mM.

[0047] For comparison, Wellbutrin® was equipotent for inhibiting DA andnoradrenaline uptake with IC₅₀ values of 1.9 and 2.2 μM, and did notinhibit serotonin uptake at 30 μM. Imipramine (a non-specific tricyclicantidepressant) inhibited NA uptake and serotonin uptake with IC₅₀values of 0.072 and 0.24 μM, respectively.

[0048] The compound of formula (I) was approximately twice as potent asWeilbutrin® as an NA inhibitor but, unlike the latter, was approximately10-fold less potent as an inhibitor of dopamine uptake. These data areconsistent with the observed noradrenergic actions of Wellbutrin® andthe racemic morpholinol metabolite of bupropion,(+/−)-(2R*,3R*)-2-(3-chlorophenyl)-3,5,5-trimethyl-2-morpholinolhydrochloride, (306U73) in vivo, at their respective anti-TBZ doses(Cooper, B. R., et al, Neuropsychopharmacology, 11: p. 133-41,1994).Behavioral and electrophysiological data suggest that the effects ofWellbutrin® are mediated by a noradrenergic mechanism (ibid).

[0049] Tetrabenazine-Induced Behavioural Depression Experiments

[0050] Tetrabenazine(TBZ)-induced behavioural depression was used as anin vivo measure of antidepressant activity. The test has been validatedwith a wide range of antidepressants, known to act through noradrenergicmechanisms (Cooper B. R. et al, “Animal models used in the prediction ofantidepressant effects in man”, J. Clin. Psychiatry 44: 63-66, 1983).Moreover, the test was also used to identify Wellbutrin® as ananti-depressant. Briefly, animals were injected with the candidate agent(p.o. or i.p.) 30 minutes before receiving an i.p. injection oftetrabenazine (35 mg/kg, as the HCl salt—prepared fresh for each use).Assessments were performed 30 minutes thereafter and included: locomotoractivity (1-4 scale); ptosis (1-4 scale) and body temperature asdescribed previously (Cooper, B. R., J. L. Howard, and F. E. Soroko,Animal models used in prediction of antidepressant effects in man(Journal of Clinical Psychiatry, 44: p. 63-6,1983). In all studies, thescientist performing the assessments was blind to the treatments. Allparameters were weighted equally to give a “lumped” score (X) throughthe following algorithm:

X=(1+Ptosis score)/(Activity score*[Temp,treated/Temp,control]

[0051] Results from the tetrabenazine-induced behavioural depressionmodel are as follows. Assessed in vivo at 25 mg/kg (ip) the compound offormula (I), the racemate, Welibutrin® and, for comparison,amitryptyline all abolished the tetrabenazine-induced behaviouraldepression. In contrast, the (−) enantiomer showed only modest activity(FIG. 1).

[0052] In the TBZ model of behavioural depression, activity resided inthe compound of formula (I). When analysed in a dose-effect study withTBZ, the activity showed a sharp increase in activity between 3 mg/kgand 6 mg/kg (ip) (FIG. 2). The compound of formula II, in comparison,did not possess dose-related activity and, at 50 mg/kg, appeared toworsen the animal's condition (FIG. 3). In FIGS. 2 and 3, AMIT (5)refers to amitryptiline dosed at 5 mg/kg and SHAM refers to a controlgroup of animals that have received no medication at all.

[0053] Since the TBZ test has been predictive of anti-depressants actingthrough noradrenergic mechanisms and the compound of formula (I) is aninhibitor of noradrenaline uptake and Wellbutrin® is metabolised to thismorpholinol in vivo, the data suggest that the anti-depressant activityof Wellbutrin® is likely to result from the effects of the compound offormula (I). (Welch, R. M., A. A. Lai, and D. H. Schroeder,Pharmacological significance of the species differences in bupropionmetabolism. Xenobiotica, 17: p. 287-98,1987).

[0054] By extension, other activities of Wellbutrin® could be attributedto the compound of formula (I). In particular, a noradrenergic mechanismis common to agents used to treat ADHD (e.g. methylphenidate andamphetamine). While the molecular mechanism for Wellbutrin's effects onsmoking cessation is less well understood, a catecholaminergic pathwayis thought to participate in the behavioural reinforcing properties ofnicotine. Wellbutrin® (and, by extension, the compound of formula (I)),by augmenting NA release into brain synapses, could mimic some of theactions of nicotine and, thus, decrease the signs associated withnicotine withdrawal. Additionally, amphetamines have been used to treatobesity. The addictive properties of amphetamine, however, preclude itsuse for most obese patients. Wellbutrin® causes weight loss and, likeamphetamine, acts through a noradrenergic mechanism. (Zarrindast, M. R.and T. Hosseini-Nia, Anorectic and behavioural effects of bupropion.General Pharmacology, 19: p. 201-4,1988 and Harto-Truax, N., et al.,Effects of Bupropion on Body Weight. Journal of Clinical Psychiatry, 44:p. 183-6,1983). However, unlike amphetamine, Wellbutrin® is notaddictive. (Lamb, R. J. and R. R. Griffiths, Self-administration inBaboons and the Discriminative Stimulus Effects in Rats of Bupropion.Nomifensine, Diclofensine and Imipramine. Psychopharmacology, 102: p.183-90,1990; Bergman, J., et al., Effects of Cocaine and Related Drugsin Nonhuman Primates. III. Self-administration by Squirrel Monkeys.Journal of Pharmacology & Experimental Therapeutics, 251: p. 150-5,1989and Johanson, C. E.; and J. E. Barrett, The Discriminative StimulusEffects of Cocaine in Pigeons. Journal of Pharmacology & ExperimentalTherapeutics, 267: p. 1-8,1993). By extension, the compound of formula(I) would also be expected to have efficacy in obesity and cocaineaddiction.

[0055] Additional studies demonstrating the use of bupropion in thetreatment of the conditions referred to above have been published asfollows: sexual dysfunction (Journal of Sex and Marital Therapy, 13(4),p 239-252, 1987), obesity (Obesity Research, 9: p 544-551, 2001),seasonal affective disorder (J. Clin. Psych., 53: p 252-255, 1992),chronic fatigue syndrome (Biol. Psychiatry, 32: p 834-838, 1992).

[0056] Safety and Toxicity

[0057] Additional dose-ranging studies were performed to determine therange of safe doses for the isomers and the racemate. Animals wereobserved for the presence of serious adverse events (e.g. seizures anddeaths) following administration of the compounds of formula I, formulaII or the racemate by the oral and intraperitoneal (i.p.) routes. Thedata are presented as Table 2.

[0058] Administered orally, at 100 mg/kg p.o., seizures were observedwith the compound of formula II and the racemate but not with thecompound of formula I. Seizures were observed in all of the animals withall 3 compounds when dosed at 300 mg/kg. Additionally, the 300 mg/kgoral dose resulted in 100 and 80% lethality for the compound of formulaII and the racemate while no deaths were observed with the compound offormula I.

[0059] Administered i.p., all of the compounds produced seizures at 100mg/kg. No deaths were observed with the compound of formula I whereasthe compound of formula II and the racemate resulted in lethality of100% and 20%, respectively. At the 300 mg/kg oral dose all of thelethality was observed for all of the compounds. TABLE 2 Effects onUptake In Vitro Compound IC50 (μM) SEM [³H]-Dopamine Uptake Bupropion1.9 0.15 Formula (I) 9.3 0.41 Formula (II) >100 [³H]-NoradrenalineUptake Bupropion 2.2 0.7 Formula (I) 1.1 0.07 Formula (II) >30Imipramine 0.072 0.020 [³H]-Serotonin Uptake Bupropion >30 Formula(I) >30 Formula (II) >100 Imipramine 0.24 0.03

[0060] TABLE 2 Adverse Events Associated with Compounds of Formula I,Formula II and the Racemate Time to Time to Dose Seizures Seizures DeathCompound Route (mg/kg) (%) (min) % Died (min) Formula I i.p. 100 1003.93 0 n/a Formula I p.o. 100 0 n/a 0 n/a Formula I i.p. 300 100 3.95100 6 Formula I p.o. 300 100 11.23 0 n/a Formula II i.p. 100 20 5 100 7Formula II p.o. 100 100 7.2 0 n/a Formula II i.p. 300 100 1.1 100 6Formula II p.o. 300 100 6.8 100 7 Racemate i.p. 100 100 3 20 14 Racematep.o. 100 100 9.2 0 n/a Racemate i.p. 300 100 3 100 3 Racemate p.o. 300100 6.8 80 7

1. A method of treatment of seasonal affective disorder in a mammalcomprising the administration to said mammal of an effective amount of(+)-(2S,3S)-2-(3-chlorophenyl)-3,5,5-trimethyl-2-morpholinol orpharmaceutically acceptable salts and solvates thereof.
 2. A method asclaimed in claim 1 comprising the administration of the compound(+)-(2S,3S)-2-(3-chlorophenyl)-3,5,5-trimethyl-2-morpholinolhydrochloride.
 3. A method as claimed in claim 2 wherein said compoundhas an optical rotation [α]²⁰ _(D) at c=0.64 in 95% EtOH of +31.9°.
 4. Amethod of treatment of chronic fatigue in a mammal comprising theadministration to said mammal of an effective amount of(+)-(2S,3S)-2-(3-chlorophenyl)-3,5,5-trimethyl-2-morpholinol orpharmaceutically acceptable salts and solvates thereof.
 5. A method asclaimed in claim 4 comprising the administration of the compound(+)-(2S,3S)-2-(3-chlorophenyl)-3,5,5-trimethyl-2-morpholinolhydrochloride.
 6. A method as claimed in claim 5 wherein said compoundhas an optical rotation [α]²⁰ _(D) at c=0.64 in 95% EtOH of +31.9°.
 7. Amethod of treatment of narcolepsy in a mammal comprising theadministration to said mammal of an effective amount of(+)-(2S,3S)-2-(3-chlorophenyl)-3,5,5-trimethyl-2-morpholinol orpharmaceutically acceptable salts and solvates thereof.
 8. A method asclaimed in claim 7 comprising the administration of the compound(+)-(2S,3S)-2-(3-chlorophenyl)-3,5,5-trimethyl-2-morpholinolhydrochloride.
 9. A method as claimed in claim 8 wherein said compoundhas an optical rotation [α]²⁰ _(D) at c=0.64 in 95% EtOH of +31.9°. 10.A method of treatment of cognitive impairment in a mammal comprising theadministration to said mammal of an effective amount of(+)-(2S,3S)-2-(3-chlorophenyl)-3,5,5-trimethyl-2-morpholinol orpharmaceutically acceptable salts and solvates thereof.
 11. A method asclaimed in claim 10 comprising the administration of the compound(+)-(2S,3S)-2-(3-chlorophenyl)-3,5,5-trimethyl-2-morpholinolhydrochloride.
 12. A method as claimed in claim 11 wherein said compoundhas an optical rotation [α]²⁰ _(D) at c=0.64 in 95% EtOH of +31.9°.